CN101096641A - Culture medium for producing winter worm summer herb mycelium - Google Patents

Culture medium for producing winter worm summer herb mycelium Download PDF

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Publication number
CN101096641A
CN101096641A CNA2007100243249A CN200710024324A CN101096641A CN 101096641 A CN101096641 A CN 101096641A CN A2007100243249 A CNA2007100243249 A CN A2007100243249A CN 200710024324 A CN200710024324 A CN 200710024324A CN 101096641 A CN101096641 A CN 101096641A
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powder
several
glucose
substratum
water
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CN101096641B (en
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李邦良
许静
王珍
袁水金
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Hangzhou Sino-American East China Pharmaceutical Jiangdong Co., Ltd.
Hangzhou Zhongmei Huadong Pharmaceutical Co Ltd
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Hangzhou Zhongmei Huadong Pharmaceutical Co Ltd
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Abstract

The invention discloses a culture medium to ferment and manufacture Cordyceps sinensis mycelium, which consists of bevel culture medium and fermentation culture medium, wherein different culture mediums are adopted in different stages to ensure plain growth of the Cordyceps sinensis and equal quality of fermented and natural Cordyceps sinensis, which modifies the product quality and improves the production with certain effect. The invention supports the character of nourishment with natural Cordyceps sinensis, which is fit for industrial manufacturing need.

Description

A kind of substratum that is used to produce Cordyceps mycelium
Technical field
Anamorph-the China pilose spore (the Latin formal name used at school is Hirsutella sinensis) that the present invention relates to a kind of Chinese Cordyceps fungus (the Latin formal name used at school is Cordyceps sinensis) adopts the production method of industrial fermentation production fermentation Cordyceps fungus powder, specifically relates to the prescription of its substratum in Industrial processes.
Background technology
Cordyceps sinensis is a kind of naturally occurring rare Chinese medicinal materials, except that containing general composition such as sugar, fat, robust fibre, also contains N.F,USP MANNITOL, ergosterol, adenosine, cordycepic acid, Cordyceps sinensis element, vitamins B 12The isoreactivity composition.Its nourishing and treatment function are on genseng, pilose antler.But natural cordyceps is long vegetative period, is distributed in high high mountain steppe area above sea level, resource scarcity, and collection is difficult for, and price is very expensive.
Zhongmei Huadong Pharmaceutical Co., Ltd. Hangzhou obtains Cordyceps mycelium and preparation " hundred make capsule " (including fermented aweto hypha body powder) thereof with artificial fermentation's method, and has recorded in " Pharmacopoeia of People's Republic of China 2005 version two ones ".
In addition, at Chinese patent " fermentation method for producing of Chinese Cordyceps sinensis fungi " (patent No. ZL97110448.4, the fermentation method for producing of a kind of Chinese Cordyceps sinensis fungi is disclosed the March 28 calendar year 2001 day for announcing), this method is bacterial classification to be implanted liquid nutrient medium place on the shaking table, temperature is below 20 ℃, cultivated 8 days, implant first class seed pot then, temperature is below 20 ℃, fermented 5-8 days, and to cultivate 10 times of expansions of base unit weight, cultivated step by step then, until reaching needed amount, go out a jar filtration and form.Carry out in the equal liquid medium within of this fermenting process, substratum is: carbon source 0.5-5%, nitrogenous source 0.2-5%, trace element, VITAMIN a little, all the other are moisture content.
This patent just discloses the prescription of liquid nutrient medium, the slant medium of bacterial classification is not disclosed, and the slant culture of bacterial classification is the starting point of whole process of production, and the slant culture that does not have a bacterial classification just can not be for fermentative production provides seed, thus the production of the Cordyceps fungus powder that can't ferment; And, natural cordyceps is as severe cold plateau species, it is grown under the environment of Plain, it is the comparison difficulty, have only the slant medium that select to be fit to the Cordyceps sinensis bacteria growing could guarantee newly to gather the successful separation of bacterial classification and constantly going down to posterity of pure Cordyceps strain, and keep its good character, thereby provide high quality seed for follow-up fermentative production, therefore, the slant culture of Cordyceps sinensis is vital in whole process of production.Slant culture belongs to solid culture, its growing environment is all different with liquid culture with nutritional requirement, the composition of slant medium is not disclosed in Chinese patent ZL97110448.4, if carry out slant culture with the liquid nutrient medium in this patent, bad phenomenon can appear in the inclined-plane growth, can't provide a favorable security for fermentation; In this external present document all not to the report of slant medium.
In addition, adopt the liquid nutrient medium among the Chinese patent ZL97110448.4 to carry out industrial production, find there is yield poorly, defective that cost is big, and the beef soup in this patent culture medium prescription etc. influenced by starting material bigger, difficult quality control etc. are not suitable for the needs of industrialized production.
The content of invention
The object of the invention is to solve deficiency such as no solid medium and liquid nutrient medium difficult quality control incompatibility suitability for industrialized production in the prior art, provides a kind of slant culture method that is used for fermented aweto hypha, for fermentation provides safeguard; Simultaneously also provide a kind of starting material nutritious, cheap, produce control easily, thereby can produce fermention medium high-quality, stay-in-grade Cordyceps fungus powder.
The invention discloses a kind of substratum that microbial fermentation is produced Cordyceps mycelium that is used for, comprise slant medium and fermention medium, it is characterized in that inclined-plane and fermentation stage adopt different substratum.
Slant medium disclosed by the invention, volume ratio (g/ml) meter comprises carbon source 1.0~8.0% by weight, nitrogenous source 1.6~8.0%, inorganic salt 0~0.1%, agar powder 0.5~1.5%, all the other are water; Preferably: carbon source 3.0~5.0%, nitrogenous source 2.5~6.0%, inorganic salt 0.02~0.06%, agar powder 0.5~1.0%, all the other are water; Wherein said carbon source is selected from several in glucose, Semen Maydis powder, murphy juice and the dextrin; Nitrogenous source is selected from yeast powder, wheat bran, dried silkworm chrysalis meal, or adds peptone again; Inorganic salt are selected from several in sodium-chlor, magnesium chloride, sal epsom, Manganous chloride tetrahydrate, Repone K, calcium chloride, potassium primary phosphate, ferrous sulfate, the manganous sulfate.
Preferred slant medium is a glucose 1.0~3.0%, Semen Maydis powder 0~2.0%, murphy juice 0~1.0%, dextrin 0~1.0%; Yeast powder 0.1~1.0%, wheat bran 0.5~2.0%, dried silkworm chrysalis meal 1.0~3.0%, peptone 0~2.0%; Sal epsom 0~0.05%, potassium primary phosphate 0~0.05%; Agar powder 0.5~1.0%, all the other are water.
Fermention medium disclosed by the invention, volume ratio (g/ml) is counted by weight, carbon source 2.0~4.0%, nitrogenous source 2.0~4.0%, inorganic salt 0~0.1%; All the other are water; Wherein said carbon source is several in glucose, corn steep liquor, the molasses; Nitrogenous source is several in dried silkworm chrysalis meal, soybean cake powder, the yeast extract paste; Inorganic salt are one or more in sodium-chlor, magnesium chloride, sal epsom, Manganous chloride tetrahydrate, Repone K, calcium chloride, potassium primary phosphate, ferrous sulfate, the manganous sulfate.
Wherein preferred fermention medium is a glucose 1.0~1.5%, corn steep liquor 0~1.0%, molasses 1.0~1.5%; Dried silkworm chrysalis meal 0.5~1.0%, soybean cake powder 1.0~2.0%, yeast extract paste 0.5~1.0%; Sal epsom 0~0.05%, potassium primary phosphate 0~0.05%, all the other are water.Preferred fermention medium is a glucose 1.0%, molasses 1.0%, and dried silkworm chrysalis meal 0.5%, soybean cake powder 1.0%, yeast extract paste 0.5%, sal epsom 0.01%, potassium primary phosphate 0.02%, all the other are water.
In industrial production, earlier with bacterial classification inoculation in slant medium, cultivated 25~35 days at 12~16 ℃, then with bacterial classification inoculation in fermentation culture based on shaking table on, temperature was cultivated 8~10 days for 12~16 ℃, be inoculated in first class seed pot then, cultivated 8~10 days for temperature 12-16 ℃, then to cultivate 8~10 times of expansions of base unit weight, fermentation step by step, until reaching required amount, carry out solid-liquid separation after going out jar, with the solid oven dry, pulverize and form.
Substratum provided by the present invention adopts different substratum at different cultivation stage, has both guaranteed that Cordyceps fungus can grow in the Plain, has guaranteed that again fermented aweto hypha body powder has the quality suitable with natural cordyceps.
The main advantage that the present invention adopts different substratum to bring is: it is to contain wheat bran in the nitrogenous source that slant medium is different from the more main of fermention medium, adopt yeast powder and dried silkworm chrysalis meal simultaneously, these three kinds of raw-material combinations, the Cordyceps fungus that has guaranteed new collection through evidence can be grown on the inclined-plane, and guaranteed to go down to posterity stable, be difficult for degenerating, for production provides guarantee.
The present invention optimizes the liquid nutrient medium among the Chinese patent ZL97110448.4, by a large amount of lab scales and pilot scale, determined the industrialized substratum of optimum, and certain effect is all arranged aspect the output improving quality product and improve, both ensured that it has the characteristics suitable with natural cordyceps nutrition, made it be suitable for the needs of industrialized production again.Fermention medium of the present invention is compared with the substratum on the Chinese patent ZL97110448.4, the environment of growing according to natural cordyceps, in order to be more suitable for the needs of industrialized production, the process modification of being undertaken by a large amount of experiments, its difference is to change glucose into by sucrose in the selection of carbon source, and this more helps the absorption of thalline; And on nitrogenous source, increased these starting material of yeast extract paste again, more help comprising the generation of main metabolites such as amino acid, adenosine, N.F,USP MANNITOL; In addition, the starting material in the substratum all are grain raw materials basically, be simple and easy to, nutritious, cheap, and through after the elaborate combination of substratum, can reduce the total amount of substratum widely, thereby reduce production costs further, need of industrial production preferably.So not only cost is low to produce Cordyceps mycelium with the present invention, is suitable for need of industrial production, and Functionality, quality and appealing design, has the nutritive value suitable with natural cordyceps.
Embodiment:
Of the present invention 1.0% murphy juice is meant: get the 10g potato, use 1000ml water, its filtrate is got in well-done filtration, is settled to 1000ml.
Embodiment 1.
In the inclined-plane, culture medium prescription is a glucose 1.5% with bacterial classification inoculation, Semen Maydis powder 1.0%, dextrin 0.5%, wheat bran 1.5%, yeast powder 0.5%, dried silkworm chrysalis meal 1.5%, peptone 1.0%; Sal epsom 0.01%, potassium primary phosphate 0.02%; Agar powder 0.8%, all the other are water, cultivate 25 days at 12 ℃; Then with bacterial classification inoculation in fermention medium, culture medium prescription is a glucose 1.0%, molasses 1.0%; Dried silkworm chrysalis meal 0.5%, soybean cake powder 1.0%, yeast extract paste 0.5%; Sal epsom 0.01%, potassium primary phosphate 0.02%, all the other are water, place on the shaking table, temperature was cultivated 8 days for 14 ℃; Be inoculated in the one grade fermemtation jar then, temperature was cultivated 8 days for 14 ℃, and to cultivate 10 times of expansions of base unit weight, fermentation step by step is until 30 tons of jars then.Carry out solid-liquid separation after going out jar, solid oven dry, pulverizing are formed.
Embodiment 2: the prescription of fermention medium is: glucose 1.5%, corn steep liquor 1.0%, molasses 1.5%; Dried silkworm chrysalis meal 1.0%, 2.0%, yeast extract paste 1.0%; Manganous sulfate 0.0125%, Repone K 0.01%, all the other are water, air flow is 5: 1.Slant medium and all the other Step By Conditions are identical with embodiment 1.
Embodiment 3: in the inclined-plane, culture medium prescription is with bacterial classification inoculation: glucose 1.0%, Semen Maydis powder 2.0%, murphy juice 1.0%, dextrin 1.0%; Wheat bran 1.5%, yeast powder 0.1%, dried silkworm chrysalis meal 3.0%; Sal epsom 0.01%, Repone K 0.01%; Agar powder 0.8%, all the other are water, fermention medium and all the other Step By Conditions are identical with embodiment 1.
Embodiment 4: the contrast experiment
Embodiment 1,2 and 3 products that obtained and product by the disclosed embodiment production of ZL97110448.4 compared (concrete data see the following form.Except that culture medium prescription and added amount difference thereof, other working conditions are equal to), the output of finding the fermented aweto hypha body that the present invention produced is higher than the output of the Cordyceps mycelium that " Chinese patent ZL97110448.4 " produced.And the measuring method of pressing " hundred make capsule " recorded in " Pharmacopoeia of People's Republic of China version in 2005 " detects it, whole standards of pharmacopoeia that meet that the present invention produced, and there is certain deviation at the cordyceps sinensis mycelium powder that " Chinese patent ZL97110448.4 " produced end with standards of pharmacopoeia.
The present invention's process is to raw-material meticulous screening, both just the added amount of fermention medium was seldom, such as carbon source and nitrogenous source all only with 2%, as embodiment 1 and embodiment 3 disclosed technical schemes, each embodiment is suitable in its output and " the ZL97110448.4 patent ", and that industrial cost has reduced is many.And if adopt with " ZL97110448.4 patent " in the amount of the suitable substratum of each embodiment, as embodiment 2 disclosed technical schemes, then output is better than it widely.
The present invention compares with the fermented aweto hypha physique amount that adopts Chinese patent ZL97110448.4 technical scheme to produce
Index " hundred make capsule " standards of pharmacopoeia Sample 1 Sample 2 Sample 3 Sample 4 Sample 5 Sample 6
Dry weight g/l / 6.7 6.3 6.5 6.6 7.9 7.0
Total amino acid (mg) Must not be less than " 60mg " 56 55 55 64 66 65
N.F,USP MANNITOL (mg) Must not be less than " 14mg " 12 11 12 15 16 16
Adenosine (mg) Must not be less than " 0.16mg " 0.15 0.14 0.13 0.20 0.24 0.22
Wherein, the fermented aweto hypha body powder of sample 1 for adopting " ZL97110448.4 patent working example 1 " to produce,
The fermented aweto hypha body powder of sample 2 for adopting " ZL97110448.4 patent working example 2 " to produce,
The fermented aweto hypha body powder of sample 3 for adopting " ZL97110448.4 patent working example 3 " to produce,
The fermented aweto hypha body powder of sample 4 for adopting this patent embodiment 1 to produce,
The fermented aweto hypha body powder of sample 5 for adopting this patent embodiment 2 to produce,
The fermented aweto hypha body powder of sample 6 for adopting this patent embodiment 3 to produce.
Wherein, the content of total amino acid, adenosine, N.F,USP MANNITOL is contained amount in every 0.2g fermented aweto hypha body powder.Can draw from above form, the present invention is used for producing, the yield height, and quality is better than ZL97110448.4 simultaneously, and meets standards of pharmacopoeia.

Claims (7)

1. one kind is used for the substratum that microbial fermentation is produced Cordyceps mycelium, comprises slant medium and fermention medium.
2. substratum according to claim 1 is characterized in that, volume ratio meter by weight, and described slant medium comprises carbon source 1.0~8.0%, nitrogenous source 1.6~8.0%, inorganic salt 0~0.1%, agar powder 0.5~1.5%, all the other are water; Wherein said carbon source is selected from several in glucose, Semen Maydis powder, murphy juice or the dextrin; Nitrogenous source is selected from several in yeast powder, dried silkworm chrysalis meal, peptone or the wheat bran; Inorganic salt are selected from several in sodium-chlor, magnesium chloride, sal epsom, Manganous chloride tetrahydrate, Repone K, calcium chloride, potassium primary phosphate, ferrous sulfate or the manganous sulfate.
3. substratum according to claim 2 is characterized in that, volume ratio meter by weight, and carbon source 3.0~5.0%, nitrogenous source 2.5~6.0%, inorganic salt 0.02~0.06%, agar powder 0.5~1.0%, all the other are water; Wherein said carbon source is selected from several in glucose, Semen Maydis powder, murphy juice and the dextrin; Nitrogenous source is selected from several in yeast powder, dried silkworm chrysalis meal, peptone and the wheat bran; Inorganic salt are selected from several in sodium-chlor, magnesium chloride, sal epsom, Manganous chloride tetrahydrate, Repone K, calcium chloride, potassium primary phosphate, ferrous sulfate, the manganous sulfate.
4. according to the described substratum of claim 3, it is characterized in that volume ratio meter by weight, glucose 1.0~3.0%, Semen Maydis powder 0~2.0%, murphy juice 0~1.0%, dextrin 0~1.0%; Yeast powder 0.1~1.0%, wheat bran 0.5~2.0%, dried silkworm chrysalis meal 1.0~%, peptone 0~2.0%; Sal epsom 0~0.05%, potassium primary phosphate 0~0.05%; Agar powder 0.5~1.0%, all the other are water.
5. substratum according to claim 1 is characterized in that, volume ratio meter by weight, and described fermention medium comprises carbon source 2.0~4.0%, nitrogenous source 2.0~4.0%, inorganic salt 0~0.1%, all the other are water; Wherein said carbon source is several in glucose, corn steep liquor, the molasses; Nitrogenous source is several in dried silkworm chrysalis meal, soybean cake powder, the yeast extract paste; Inorganic salt are one or more in sodium-chlor, magnesium chloride, sal epsom, Manganous chloride tetrahydrate, Repone K, calcium chloride, potassium primary phosphate, ferrous sulfate, the manganous sulfate.
6. substratum according to claim 5 is characterized in that, volume ratio meter by weight, described glucose 1.0~1.5%, corn steep liquor 0~1.0%, molasses 1.0~1.5%; Dried silkworm chrysalis meal 0.5~1.0%, soybean cake powder 1.0~2.0%, yeast extract paste 0.5~1.0%; Sal epsom 0~0.05%, potassium primary phosphate 0~0.05%, all the other are water.
7. substratum according to claim 6 is characterized in that, volume ratio meter by weight, described glucose 1.0%, molasses 1.0%, dried silkworm chrysalis meal 0.5%, soybean cake powder 1.0%, yeast extract paste 0.5%, sal epsom 0.01%, potassium primary phosphate 0.02%, all the other are water.
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Cited By (14)

* Cited by examiner, † Cited by third party
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CN101791329A (en) * 2010-03-30 2010-08-04 天津科技大学 Astragalus waste cordyceps sinensis fermentation technique and application thereof
CN102220249A (en) * 2011-06-13 2011-10-19 杭州柯氏生物科技有限公司 Method for producing Hirsutella sinensis
CN102936609A (en) * 2012-11-05 2013-02-20 南京农业大学 Preparation method of swift moth paecilomyces varioti extracellular acid glycoprotein
CN103396193A (en) * 2013-08-08 2013-11-20 董亚贤 Cordycepsmilitaris strain liquid culture medium and preparation method therefor
CN103621315A (en) * 2013-12-12 2014-03-12 福建农林大学 Method for cultivating cordyceps militaris by using Chinese herbal medicine nutrient solution
CN103828597A (en) * 2013-07-01 2014-06-04 鲁东大学 Method for making pleurotus citrinopileatus liquid spawn
CN103988712A (en) * 2014-05-29 2014-08-20 熊艳 High-yield polysaccharide cordyceps militaris cultivation method
CN105112298A (en) * 2015-08-26 2015-12-02 中国科学院微生物研究所 Trichoderma voglmayrii producing mycelium cultivation medium, method for preparing same and application
CN106497795A (en) * 2016-10-12 2017-03-15 锬酃藏虫草生物科技(深圳)有限公司 A kind of Cordyceps funguss seed culture medium containing Fructus Lycii and its application
CN108384726A (en) * 2018-04-25 2018-08-10 杭州中美华东制药有限公司 Aweto mycelium and its fermentation medium and fermentation process
CN109355204A (en) * 2018-11-28 2019-02-19 马兰 A kind of method of fermenting and producing cordyceps sinensis mycelium powder
CN109370914A (en) * 2018-11-28 2019-02-22 马兰 A kind of cordyceps sinensis mycelium powder and its production method
WO2019223287A1 (en) * 2018-05-25 2019-11-28 江西国药有限责任公司 Culture medium for paecilomyces hepiali cs-4 and preparation method therefor
CN110628664A (en) * 2019-08-15 2019-12-31 华中农业大学 Pseudomonas eastern China for preventing and treating root-knot nematode as well as preparation method and application thereof

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101791329A (en) * 2010-03-30 2010-08-04 天津科技大学 Astragalus waste cordyceps sinensis fermentation technique and application thereof
CN102220249A (en) * 2011-06-13 2011-10-19 杭州柯氏生物科技有限公司 Method for producing Hirsutella sinensis
CN102936609B (en) * 2012-11-05 2014-12-17 南京农业大学 Preparation method of swift moth paecilomyces varioti extracellular acid glycoprotein
CN102936609A (en) * 2012-11-05 2013-02-20 南京农业大学 Preparation method of swift moth paecilomyces varioti extracellular acid glycoprotein
CN103828597A (en) * 2013-07-01 2014-06-04 鲁东大学 Method for making pleurotus citrinopileatus liquid spawn
CN103396193A (en) * 2013-08-08 2013-11-20 董亚贤 Cordycepsmilitaris strain liquid culture medium and preparation method therefor
CN103621315B (en) * 2013-12-12 2015-02-04 福建农林大学 Method for cultivating cordyceps militaris by using Chinese herbal medicine nutrient solution
CN103621315A (en) * 2013-12-12 2014-03-12 福建农林大学 Method for cultivating cordyceps militaris by using Chinese herbal medicine nutrient solution
CN103988712A (en) * 2014-05-29 2014-08-20 熊艳 High-yield polysaccharide cordyceps militaris cultivation method
CN105112298A (en) * 2015-08-26 2015-12-02 中国科学院微生物研究所 Trichoderma voglmayrii producing mycelium cultivation medium, method for preparing same and application
CN106497795A (en) * 2016-10-12 2017-03-15 锬酃藏虫草生物科技(深圳)有限公司 A kind of Cordyceps funguss seed culture medium containing Fructus Lycii and its application
CN108384726A (en) * 2018-04-25 2018-08-10 杭州中美华东制药有限公司 Aweto mycelium and its fermentation medium and fermentation process
WO2019223287A1 (en) * 2018-05-25 2019-11-28 江西国药有限责任公司 Culture medium for paecilomyces hepiali cs-4 and preparation method therefor
CN109355204A (en) * 2018-11-28 2019-02-19 马兰 A kind of method of fermenting and producing cordyceps sinensis mycelium powder
CN109370914A (en) * 2018-11-28 2019-02-22 马兰 A kind of cordyceps sinensis mycelium powder and its production method
CN109370914B (en) * 2018-11-28 2021-07-27 马兰 Cordyceps sinensis mycelium powder and production method thereof
CN110628664A (en) * 2019-08-15 2019-12-31 华中农业大学 Pseudomonas eastern China for preventing and treating root-knot nematode as well as preparation method and application thereof
CN110628664B (en) * 2019-08-15 2021-03-16 华中农业大学 Pseudomonas eastern China for preventing and treating root-knot nematode as well as preparation method and application thereof

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Patentee after: Zhongmei Huadong Pharmaceutical Co., Ltd., Hangzhou

Address before: Hangzhou City, Zhejiang province 310011 Moganshan Road No. 866

Co-patentee before: Huadong medicine (Hangzhou) bailing biological science and Technology Co Ltd

Patentee before: Zhongmei Huadong Pharmaceutical Co., Ltd., Hangzhou